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Necrostatin-1

別名: Nec-1

Necrostatin-1 (Nec-1)是一種特異性RIP1 (RIPK1)抑制劑，抑制TNF-α誘導的細胞壞死，在293T細胞中EC50為490 nM。Necrostatin-1也可抑制 IDO、細胞自噬和凋亡。

Necrostatin-1 Chemical Structure

Necrostatin-1 Chemical Structure

CAS: 4311-88-0

規(guī)格 價格 庫存 購買數(shù)量
10mM (1mL in DMSO) 1054.09 現(xiàn)貨
10mg 814.14 現(xiàn)貨
100mg 2224.79 現(xiàn)貨
1g 7944.3 現(xiàn)貨
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Necrostatin-1相關(guān)產(chǎn)品

細胞實驗數(shù)據(jù)示例

細胞系 實驗類型 給藥濃度 孵育時間 活性描述 文獻信息
L929 Growth Inhibition Assay 2/5?μg/ml? 24?h reverses the cell growth inhibition and cell death induced by TNFα alone as well as TNFα?+?zVAD 23941769
L929 Function Assay 2?μg/ml? 24?h promots caspase-6 (p20) activity and procaspase-6 cleavage 23941769
L929 Function Assay 5?μg/ml 24?h blocks zVAD induced necroptosis and autophagy 23941769
C6 Cell Viability Assay 1 mmol/L 3 h attenuates Shikonin induced glioma cell death 23840441
U87 Cell Viability Assay 1 mmol/L 3 h attenuates Shikonin induced glioma cell death 23840441
C6 Cytotoxicity Assay 1 mmol/L 3 h blocks shikonin induced necrosis 23840441
U87 Cytotoxicity Assay 1 mmol/L 3 h blocks shikonin induced necrosis 23840441
C6 Function Assay 1 mmol/L 1.5-3 h suppresses the expression of RIP-1 caused by shikonin 23840441
U87 Function Assay 1 mmol/L 1.5-3 h suppresses the expression of RIP-1 caused by shikonin 23840441
TE671 Cell Viability Assay 40?μg/ml? 24 h rescues GX15-070-induced loss of cell viability 23744296
RMS13 Cell Viability Assay 40?μg/ml? 24 h rescues GX15-070-induced loss of cell viability 23744296
MEFs Cytotoxicity Assay 2/6/20 μM 18 h inhibits TNFα-induced cell death in RelA KO MEFs 23727581
MEFs Function Assay 20?μM 1/2/4 h suppresses TNFα-induced RIPK1 phosphorylation 23727581
ΔN-Karpas 299? Cytotoxicity Assay 20?μM 16 h inhibits CD30-induced cell death 23545938
MM.1S? Cytotoxicity Assay 90 μM 1 h blocks BAY 11-7082 induced rapid cell swelling 23527154
KMS-12-BM Cytotoxicity Assay 90 μM 1 h blocks BAY 11-7082 induced rapid cell swelling 23527154
HT-22? Cell Viability Assay 10?μM 12?h protects against glutamate-induced cell death 23307752
HT-22? Function Assay 25?μM 0–30?min inhibits ERK Activation induced by glutamate 23307752
NIH3T3? Function Assay 10/50 μM 1/3 h ameliorates TNFα-driven complex formation 23261677
SH-EP Apoptosis Assay 10?μM? 72?h? inhibits IAP inhibitor- and Lexatumumab-induced apoptosis 22890322
HL60 Apoptosis Assay 60 μM 12 h enhances shikonin-induced apoptosis 22837689
HL60/Adr Apoptosis Assay 60 μM 12 h enhances shikonin-induced apoptosis 22837689
K562 Apoptosis Assay 60 μM 12 h enhances shikonin-induced apoptosis 22837689
K562/Adr? Apoptosis Assay 60 μM 12 h enhances shikonin-induced apoptosis 22837689
HL60 Function Assay 60 μM 12 h augments the caspase-3 activity 22837689
HL60/Adr Function Assay 60 μM 12 h augments the caspase-3 activity 22837689
K562 Function Assay 60 μM 12 h augments the caspase-3 activity 22837689
K562/Adr? Function Assay 60 μM 12 h augments the caspase-3 activity 22837689
HL60 Function Assay 60 μM 12 h increases the activity of caspases, caspase 8 and 9 22837689
HL60/Adr Function Assay 60 μM 12 h increases the activity of caspases, caspase 8 and 9 22837689
K562 Function Assay 60 μM 12 h increases the activity of caspases, caspase 8 and 9 22837689
K562/Adr? Function Assay 60 μM 12 h increases the activity of caspases, caspase 8 and 9 22837689
L929sA Apoptosis Assay 10 μM 1 h inhibits the apoptotic response to TNF 22362767
L929sA Apoptosis Assay 10 μM 1 h rescues cells expressing RIPK1ΔID from TNF-induced apoptosis 22362767
L929sA Apoptosis Assay 10 μM 1 h abrogates the interaction of caspase-8 with FADD 22362767
TPC-1 Cell Viability Assay 100 μM 24 h increases cellular survival 22136818
8505c Cell Viability Assay 100 μM 24 h increases cellular survival 22136818
SW13 Cell Viability Assay 100 μM 24 h increases cellular survival 22136818
Jurkat? Cytotoxicity Assay 50/ 100/200?μm 1/3 h reduces?Naegleria fowleri-induced cytotoxicity 21535020
Jurkat? Function Assay 200?μm 30 min reduces?Naegleria fowleri-induced reactive oxygen species (ROS) generation 21535020
HT-22 Cytotoxicity Assay 10 μM 12 h protects against cell death induced by 5?mmol/L glutamate? 17760869
L929 Function Assay 2/5?μg/ml? 24?h reversed the autophagy induced by TNFα alone as well as TNFα?+?zVAD 23941769
NRK-52E? Cell Viability Assay 20 μM 24 h inhibits increased Drp1 protein expression after TNF-α Stimulation and ATP Depletion 24351845
NRK-52E? Cell Viability Assay 20 μM 24 h increases cell viability after TNF-α Stimulation and ATP Depletion 24351845
NRK-52E? Cell Viability Assay 20 μM 24 h protects cells from cell death caused by ischemia injury 24351845
AGS Cell Viability Assay 60?μm 1?h prevents shikonin-induced cell death 24463199
L-540? Cell Viability Assay 60?μm 1?h reduces the Givinostat/Sorafenib-induced cell death 24561519
L-540? Function Assay 60?μm 1?h prevents the mitochondrial membrane depolarization 24561519
L-540? Function Assay 60?μm 1?h prevents the generation of ROS 24561519
SK-Hep1 Function Assay 60?μM? 18?h blocks?β-lapachone-mediated PAR accumulation and AIF translocation to the cytosol? 24832602
SK-Hep1 Function Assay 60?μM? 18?h inhibits β-Lapachone-induced leakage of HMGB-1? 24832602
SK-Hep1 Function Assay 60?μM? 18?h blocks?β-lapachone-induced morphological change, cell death and PI uptake 24832602
Huh7 Cell Viability Assay 50 μM 24/48 h prevents cell death of rAdHCV co-infected Huh7 cells 24973240
L929 Cell Viability Assay 30?μM 1?h inhibits TNF-α-induced cleavage of Topo I 25095742
L929 Cell Viability Assay 30?μM 1?h inhibits TNF-α-induced loss of cell viability 25095742
L929-A Function Assay 50?μM? 12 h inhibits the TNFα-induced loss of mitochondrial membrane permeability 25398540
L929 Function Assay 50?μM? 12 h inhibits TNFα-induced Bid cleavage 25398540
L929-N? Function Assay 50?μM? 12 h blocks the cleavage of Caspase-3 and PARP 25398540
L929-A? Function Assay 50?μM? 12 h blocks the cleavage of Caspase-3 and PARP 25398540
L929-N? Cell Viability Assay 50?μM? 24 h blocks TNFα-induced cell death 25398540
L929-A? Cell Viability Assay 50?μM? 24 h blocks TNFα-induced cell death 25398540
KMS-12-PE? Cell Viability Assay 60 μM 5 h inhibits SHK-induced cell death 25530098
SGC-7901 Cell Viability Assay 30?μM 1?h suppresses oxaliplatin-mediated cell death 25767076
BxPC-3 Function Assay 20 μM 24 h decreases the early necrotic cells 26000607
MiaPaCa-2 Function Assay 20 μM 24 h decreases the early necrotic cells 26000607
NCI-H28 Cell Viability Assay 10?μM 24?h prevents DAPE-induced reduction of NCI-H28 cell viability? 26004138
BMDM? Function Assay 10?μM 30?min protects cells from TAKI-induced LDH release 26381601
MEFs? Cell Viability Assay 10?μM 48 h inhibits zVAD-promoted death of CNOT3-depleted MEFs 26437789
A549 Cell Viability Assay 50?μM 24?h inhibits MMS-induced cell death 26472723
Jurkat T Necroptosis assay 30 uM 24 hrs Inhibition of necroptosis in TNF-alpha-induced human Jurkat T cells assessed as cell viability at 30 uM after 24 hrs 18467094
L929 Necroptosis assay 30 uM 24 hrs Inhibition of necroptosis in zVAD-induced mouse L929 cells assessed as cell viability at 30 uM after 24 hrs 18467094
L929 Necroptosis assay 30 uM 24 hrs Inhibition of necroptosis in TNF-alpha-induced mouse L929 cells assessed as cell viability at 30 uM after 24 hrs 18467094
Jurkat Cytoprotective assay 30 uM 1 hr Cytoprotective activity against FasL-induced necroptosis in human Jurkat cells assessed as increase in cell viability at 30 uM incubated for 1 hr followed by FasL stimulation measured after 20 hrs by Alamar blue assay 29541357
Jurkat Cytoprotective assay 30 uM 1 hr Cytoprotective activity against CHX-induced necroptosis in human Jurkat cells assessed as increase in cell viability at 30 uM incubated for 1 hr followed by CHX stimulation by Alamar blue assay 29541357
Jurkat Cytoprotective assay 30 uM 1 hr Cytoprotective activity against Z-VAD-induced necroptosis in human Jurkat cells assessed as increase in cell viability at 30 uM incubated for 1 hr followed by Z-VAD stimulation by Alamar blue assay 29541357
Jurkat Cytoprotective assay 30 uM 1 hr Cytoprotective activity against FasL-induced necroptosis in human Jurkat cells assessed as increase in cell viability at 30 uM incubated for 1 hr followed by FasL stimulation measured after 20 hrs by phase contrast microscopy 29541357
Jurkat Cytoprotective assay 30 uM 1 hr Cytoprotective activity against CHX-induced necroptosis in human Jurkat cells assessed as increase in cell viability at 30 uM incubated for 1 hr followed by CHX stimulation by phase contrast microscopy 29541357
Jurkat Cytoprotective assay 30 uM 1 hr Cytoprotective activity against Z-VAD-induced necroptosis in human Jurkat cells assessed as increase in cell viability at 30 uM incubated for 1 hr followed by Z-VAD stimulation by phase contrast microscopy 29541357
OHC Function Assay 300?μM increases the number of apoptotic OHCs without altering the levels of CC8 after noise exposure 24874734
OHC Function Assay 300?μM diminishes noise-induced AMPK activation 24874734
OHC Function Assay 300?μM results in a reduction of noise-induced RIP1 and RIP3 immunofluorescence 24874734
OHC Function Assay 300?μM decreases noise-induced swollen nuclei? 24874734
OHC Function Assay 300?μM increases noise-induced condensed nuclei 24874734
Sf9 Function assay 30 mins Inhibition of recombinant human GST-fused RIPK1 (1 to 497 residues) expressed in baculovirus infected insect Sf9 cells in presence of 32P-gamma-ATP after 30 mins by autoradiogram-based Western blot method, IC50 = 0.182 μM. 28280261
3T3 Cell death assay 24 hrs Inhibition of death receptor signaling mediated necrotic cell death in mouse 3T3 cells assessed as cell viability after 24 hrs by ATP based viability assay in presence of TNFalpha and zVAD.fmk 16408008
3T3 Cell death assay 24 hrs Inhibition of death receptor signaling mediated necrotic cell death in mouse 3T3 cells assessed as cell viability after 24 hrs by ATP based viability assay in presence of FasL and zVAD.fmk 16408008
MEF Cell death assay 16 hrs Inhibition of death receptor signaling mediated necrotic cell death in SV40 transformed mouse MEF cells assessed as cell viability after 16 hrs by ATP based viability assay in presence of TNFalpha, CHX and zVAD.fmk 16408008
L929 Cell death assay 24 hrs Inhibition of death receptor signaling mediated necrotic cell death in mouse L929 cells assessed as cell viability after 24 hrs by ATP based viability assay in presence of TNFalpha 16408008
U937 Cell death assay 48 hrs Inhibition of death receptor signaling mediated necroptotic cell death in human U937 cells assessed as cell viability after 48 hrs by ATP based viability assay in presence of TNFalpha and zVAD-fmk 16408008
3T3 Cell death assay 24 hrs Inhibition of death receptor signaling mediated necroptotic cell death in mouse 3T3 cells assessed as cell viability after 24 hrs by ATP based viability assay in presence of TNFalpha and zVAD-fmk 16408008
Jurkat Cell death assay 24 hrs Inhibition of death receptor signaling mediated necroptotic cell death in human Jurkat cells deficient in FADD assessed as decreased levels of PE-conjugated LC3-II (autophagy marker) after 24 hrs by Western blot method in presence of TNFalpha 16408008
L929 Cell death assay 24 hrs Inhibition of death receptor signaling mediated necroptotic cell death in mouse L929 cells assessed as decreased levels of PE-conjugated autophagy marker LC3-II after 24 hrs by Western blot method in presence of TNFalpha 16408008
3T3 Cell death assay 24 hrs Inhibition of death receptor signaling mediated necroptotic cell death in mouse 3T3 cells assessed as decreased levels of PE-conjugated autophagy marker LC3-II after 24 hrs by Western blot method in presence of TNFalpha and zVAD-fmk 16408008
3T3 Cell death assay 24 hrs Inhibition of death receptor signaling mediated necroptotic cell death in mouse 3T3 cells assessed as decreased levels of PE-conjugated autophagy marker LC3-II after 24 hrs by Western blot method in presence of FasL and zVAD-fmk 16408008
3T3 Cell death assay 24 hrs Inhibition of death receptor signaling mediated necroptotic cell death in mouse 3T3 cells assessed as decreased levels of PE-conjugated autophagy marker LC3-II after 24 hrs by Western blot method in presence of rapamycin 16408008
Jurkat Cell death assay 48 hrs Inhibition of death receptor signaling mediated necroptotic cell death in human Jurkat cells deficient in FADD and expressing FKBP12-based dimerization domain assessed as cell viability after 48 hrs by FACS in presence of AP1510, zVAD-fmk 16408008
Jurkat Cell death assay 48 hrs Inhibition of death receptor signaling mediated necroptotic cell death in human Jurkat cells deficient in FADD and expressing RIP kinase assessed as cell viability after 48 hrs by FACS in presence of AP1510, zVAD-fmk 16408008
Jurkat Cell death assay 48 hrs Inhibition of death receptor signaling mediated necroptotic cell death in human Jurkat cells deficient in FADD and expressing RIP K45M mutant assessed as cell viability after 48 hrs by FACS in presence of AP1510, zVAD-fmk 16408008
Jurkat Cell death assay 48 hrs Inhibition of death receptor signaling mediated necroptotic cell death in human Jurkat cells deficient in FADD and expressing RIP kinase domain assessed as cell viability after 48 hrs by FACS in presence of AP1510, zVAD-fmk 16408008
Jurkat Cell death assay 48 hrs Inhibition of death receptor signaling mediated necroptotic cell death in human Jurkat cells deficient in FADD and expressing FKBP12-based dimerization domain assessed as cell viability after 48 hrs by FACS in presence of AP1510 16408008
Jurkat Cell death assay 48 hrs Inhibition of death receptor signaling mediated necroptotic cell death in human Jurkat cells deficient in FADD and expressing RIP kinase assessed as cell viability after 48 hrs by FACS in presence of AP1510 16408008
Jurkat Cell death assay 48 hrs Inhibition of death receptor signaling mediated necroptotic cell death in human Jurkat cells deficient in FADD and expressing RIP K45M mutant assessed as cell viability after 48 hrs by FACS in presence of AP1510 16408008
Jurkat Cell death assay 48 hrs Inhibition of death receptor signaling mediated necroptotic cell death in human Jurkat cells deficient in FADD and expressing RIP kinase domain assessed as cell viability after 48 hrs by FACS in presence of AP1510 16408008
Jurkat T Necroptosis assay Inhibition of TNF-alpha-induced necroptosis in FADD-deficient human Jurkat T cells, EC50 = 0.05 μM. 18467094
Jurkat Function assay Inhibition of endogenous RIP1 autophosphorylation in human Jurkat cells, EC50 = 0.182 μM. 18408713
Jurkat T Necroptosis assay Effective concentration required for inhibition of necroptosis in FADD deficient Jurkat T cells treated with TNF-alpha, EC50 = 0.49 μM. 16153840
Jurkat Necroptosis assay Inhibition of cellular necroptosis in TNFalpha treated FADD deficient human Jurkat cells, EC50 = 0.49 μM. 18408713
IEC18 Cell death assay Inhibition of death receptor signaling mediated necrotic cell death in rat IEC18 cells assessed as cell viability in presence of TNFalpha and zVAD.fmk 16408008
HL60 Cell death assay Inhibition of death receptor signaling mediated necrotic cell death in human HL60 cells assessed as cell viability in presence of TNFalpha and zVAD.fmk 16408008
Jurkat Necrosis assay Inhibition of necrosis in human Jurkat cells assessed as nuclear condensation by bright field microscopy in presence of FasL, CHX and zVAD-fmk 16408008
Jurkat Necrosis assay Inhibition of necrosis in human Jurkat cells assessed as organelle swelling by bright field microscopy in presence of FasL, CHX and zVAD-fmk 16408008
Jurkat Necrosis assay Inhibition of necrosis in human Jurkat cells assessed as early loss of plasma membrane integrity by bright field microscopy in presence of FasL, CHX and zVAD-fmk 16408008
Jurkat Necrosis assay Inhibition of necrosis in human Jurkat cells assessed as appearance of translucent cytosol in presence of FasL, CHX and zVAD-fmk 16408008
Jurkat Necrosis assay Inhibition of necrosis in human Jurkat cells deficient in FADD assessed as inhibition of nuclear condensation by bright field microscopy in presence of TNFalpha 16408008
Jurkat Necrosis assay Inhibition of necrosis in human Jurkat cells deficient in FADD assessed as inhibition of organelle swelling by bright field microscopy in presence of TNFalpha 16408008
Jurkat Necrosis assay Inhibition of necrosis in human Jurkat cells deficient in FADD assessed as inhibition of early loss of plasma membrane integrity by bright field microscopy in presence of TNFalpha 16408008
Jurkat Necrosis assay Inhibition of necrosis in human Jurkat cells deficient in FADD assessed as inhibition of appearance of translucent cytosol in presence of TNFalpha 16408008
Sf9 Function assay Inhibition of human RIP1 K45M mutant autophosphorylation expressed in Sf9 cells 18408713
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生物活性

產(chǎn)品描述 Necrostatin-1 (Nec-1)是一種特異性RIP1 (RIPK1)抑制劑，抑制TNF-α誘導的細胞壞死，在293T細胞中EC50為490 nM。Necrostatin-1也可抑制 IDO、細胞自噬和凋亡。
特性 Necrostatin-1是抑制細胞壞死的有力工具。
靶點
IDO [1] RIP1 [1]
(293T cells)
490 nM(EC50)
體外研究(In Vitro)
體外研究活性

Necrostatin-1 (1-100 μM) 抑制過表達和內(nèi)源性的RIP1發(fā)生自磷酸化。RIP1是初級細胞靶點，負責Necrostatin-1的抗細胞壞死活性。[1]

Necrostatin-1有效抑制多種類型細胞觸發(fā)的壞死性細胞死亡。Necrostatin-1作為細胞壞死的小分子抑制劑, 作用于jurkat細胞，抑制RIP激酶的誘導細胞壞死，抑制TNF-α誘導的細胞壞死，EC50為490 nM。[2]
激酶實驗 RIP1 激酶檢測
RIP1 的磷酸化需要其激酶活性。FLAG標記的野生型(WT)或RIP1(K45M) 突變體失活激酶的表達結(jié)構(gòu)轉(zhuǎn)染到293T細胞中，在有[γ-32P]ATP存在時，RIP1激酶實驗在30°C下進行30分鐘。樣品進行SDS-PAGE，通過放射自顯影可觀察到RIP1帶。對放射性帶的相對強度進行量化，并顯示比率。在激酶反應的同時，珠樣本使用anti-RIP1抗體進行Western Blot分析，確保與激酶反應中等量的蛋白。
細胞實驗 細胞系 Jurkat, BALB/c 3T3, SV40-轉(zhuǎn)化的MEF, L929
濃度 0.01-100 μM
孵育時間 --
方法

細胞接種在96孔板中（白色板進行發(fā)光檢測;黑色板進行熒光檢測;空白板進行MTT實驗）貼壁細胞按每孔5000-10000個細胞的密度接種，懸浮細胞按每孔20,000-50,000個細胞的密度接種，孔中含100 μl合適的無酚紅培養(yǎng)基。溫育后，使用如下方法之一測定細胞存活率。ATP 實驗中，使用購買的發(fā)光試劑盒，并使用Wallac Victor II酶標儀分析發(fā)光值。Sytox實驗中,細胞與 1 μM Sytox Green試劑在37°C下溫育30分鐘,然后進行熒光讀數(shù)。隨后，增加5 μl 20% Triton X-100溶液到每孔中，產(chǎn)生最大溶解，細胞37°C下溫育1小時，然后進行二次讀數(shù)。Triton處理前和后，計算值的比率。MTT實驗，使用CellTiter 96 AQueous 非放射性細胞增殖檢測試劑盒。PI排除實驗中, 加入2 μg/ml PI 到培養(yǎng)基中，立即使用FACSCalibur分析樣品。PI-膜聯(lián)蛋白V 實驗中，使用ApoAlert Annexin V-EGFP 凋亡試劑盒。進行DioC6染色, 細胞與40 nM DiOC6 在37°C下溫育30分鐘, 洗滌一次，使用FACSCalibur分析。ROS分析中, 細胞與5 μM Dihydroethidium在37°C下溫育30分鐘, 洗滌一次，使用FACSCalibur分析。
實驗圖片 檢測方法 檢測指標 實驗圖片 PMID
Immunofluorescence RIP1 / RIP3 30462730

化學信息&溶解度

分子量 259.33 分子式

C13H13N3OS
CAS號 4311-88-0 SDF Download Necrostatin-1 SDF
Smiles CN1C(=O)C(NC1=S)CC2=CNC3=CC=CC=C32
儲存條件(自收到貨起)

體外溶解度
批次：

DMSO : 57 mg/mL ( (219.79 mM) ；DMSO吸濕會降低化合物溶解度，請使用新開封DMSO)

Water : Insoluble

Ethanol : Insoluble

摩爾濃度計算器

體內(nèi)溶解配方
批次：

現(xiàn)配現(xiàn)用，請按從左到右的順序依次添加，澄清后再加入下一溶劑

 動物體內(nèi)配方計算器

實驗計算

摩爾濃度計算器

質(zhì)量 濃度 體積 分子量

動物體內(nèi)配方計算器（澄清溶液）

第一步：請輸入基本實驗信息（考慮到實驗過程中的損耗，建議多配一只動物的藥量）

mg/kg g μL 只

第二步：請輸入動物體內(nèi)配方組成（配方適用于不溶于水的藥物；不同批次藥物配方比例不同，請聯(lián)系Selleck為您提供正確的澄清溶液配方）

% DMSO % % Tween 80 % ddH2O
%DMSO %

計算結(jié)果：

工作液濃度： mg/ml；

DMSO母液配制方法： mg 藥物溶于μL DMSO溶液（母液濃度mg/mL，注：如該濃度超過該批次藥物DMSO溶解度，請先聯(lián)系Selleck）；

體內(nèi)配方配制方法：取μL DMSO母液，加入μL PEG300，混勻澄清后加入μL Tween 80，混勻澄清后加入μL ddH2O，混勻澄清。

體內(nèi)配方配制方法：取μL DMSO母液，加入μL Corn oil，混勻澄清。

注意：1. 首先保證母液是澄清的；
2.一定要按照順序依次將溶劑加入，進行下一步操作之前必須保證上一步操作得到的是澄清的溶液，可采用渦旋、超聲或水浴加熱等物理方法助溶。

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